How osteons form: A quantitative hypothesis-testing analysis of cortical pore filling and wall asymmetry.

Osteon morphology provides valuable information about the interplay between different processes involved in bone remodelling. The correct quantitative interpretation of these morphological features is challenging due to the complexity of interactions between osteoblast behaviour, and the evolving geometry of cortical pores during pore closing. We present a combined experimental and mathematical modelling study to provide insights into bone formation mechanisms during cortical bone remodelling based on histological cross-sections of quiescent human osteons and hypothesis-testing analyses. We introduce wall thickness asymmetry as a measure of the local asymmetry of bone formation within an osteon and examine the frequency distribution of wall thickness asymmetry in cortical osteons from human iliac crest bone samples from women 16-78 years old. Our measurements show that most osteons possess some degree of asymmetry, and that the average degree of osteon asymmetry in cortical bone evolves with age. We then propose a comprehensive mathematical model of cortical pore filling that includes osteoblast secretory activity, osteoblast elimination, osteoblast embedment as osteocytes, and osteoblast crowding and redistribution along the bone surface. The mathematical model is first calibrated to symmetric osteon data, and then used to test three mechanisms of asymmetric wall formation against osteon data: (i) delays in the onset of infilling around the cement line; (ii) heterogeneous osteoblastogenesis around the bone perimeter; and (iii) heterogeneous osteoblast secretory rate around the bone perimeter. Our results suggest that wall thickness asymmetry due to off-centred Haversian pores within osteons, and that nonuniform lamellar thicknesses within osteons are important morphological features that can indicate the prevalence of specific asymmetry-generating mechanisms. This has significant implications for the study of disruptions of bone formation as it could indicate what biological bone formation processes may become disrupted with age or disease.

Human femur morphology and histology variation with ancestry and behaviour in an ancient sample from Vietnam.

BACKGROUND: There is a genetic component to the minimum effective strain (MES)-a threshold which determines when bone will adapt to function-which suggests ancestry should play a role in bone (re)modelling. Further elucidating this is difficult in living human populations because of the high global genetic admixture. We examined femora from an anthropological skeletal assemblage (Mán Bac, Vietnam) representing distinct ancestral groups. We tested whether femur morphological and histological markers of modelling and remodelling differed between ancestries despite their similar lifestyles. METHODS: Static histomorphometry data collected from subperiosteal cortical bone of the femoral midshaft, and gross morphometric measures of femur robusticity, were studied in 17 individuals from the Mán Bac collection dated to 1906-1523 cal. BC. This assemblage represents agricultural migrants with affinity to East Asian groups, who integrated with the local hunter-gatherers with affinity to Australo-Papuan groups during the mid-Holocene. Femur robusticity and histology data were compared between groups of 'Migrant' (n = 8), 'Admixed' (n = 4), and 'Local' (n = 5). RESULTS: Local individuals had more robust femoral diaphyses with greater secondary osteon densities, and relatively large secondary osteon and Haversian canal parameters than the migrants. The Migrant group showed gracile femoral shafts with the least dense bone made up of small secondary osteons and Haversian canals. The Admixed individuals fell between the Migrant and Local categories in terms of their femoral data. However, we also found that measures of how densely bone is remodelled per unit area were in a tight range across all three ancestries. CONCLUSIONS: Bone modelling and remodelling markers varied with ancestral histories in our sample. This suggests that there is an ancestry related predisposition to bone optimising its metabolic expenditure likely in relation to the MES. Our results stress the need to incorporate population genetic history into hierarchical bone analyses. Understanding ancestry effects on bone morphology has implications for interpreting biomechanical loading history in past and modern human populations.

An examination of histomorphometric relationships in the anterior and posterior human femoral cortex.

INTRODUCTION: Static cortical bone histomorphometry utilised in forensic age-at-death estimation generally examines only the anterior femoral mid-shaft, as biomechanical strain at the posterior femur is thought to result in increased bone remodelling, osteon density and adversely affect age-at-death estimates. As osteon density increases there is a corresponding decrease in geometric variables, such as osteon area and Haversian canal diameter. The present study tests whether the inverse relationship between osteon density and osteon geometry is reflected in a modern documented Australian sample, and if this relationship differs between the anterior and posterior femoral mid-shaft. MATERIALS AND METHODS: The study sample comprises 215 femoral microradiographs (117♂ 98♀) of recorded age (18‒97 years) from the Melbourne Femur Reference Collection (MFRC). The following variables were measured in Image J across six 1 mm2 regions of interest (ROIs) from the anterior and posterior; mean intact and fragmentary secondary osteon count, osteon population density, osteon and Haversian canal area, perimeter, and diameter. RESULTS: Osteon area was positively correlated with Haversian canal size and shape metrics, and negatively correlated with osteon density. Chronological age was significantly correlated with most variables. There were significant between-group effects between the youngest (18‒34 years) and all other age groups (35‒49, 50-74 and 75 + years) for both regions. CONCLUSION: Our findings support an increased rate of remodelling associated with decreases in osteon geometry in the anterior and posterior femur. Future studies should examine static osteon histomorphometry using anterior and posterior measurements in larger samples of documented age and sex.

Preliminary paleohistological observations of the StW 573 ('Little Foot') skull.

Numerous aspects of early hominin biology remain debated or simply unknown. However, recent developments in high-resolution imaging techniques have opened new avenues in the field of paleoanthropology. More specifically, X-ray synchrotron-based analytical imaging techniques have the potential to provide crucial details on the ontogeny, physiology, biomechanics, and biological identity of fossil specimens. Here we present preliminary results of our X-ray synchrotron-based investigation of the skull of the 3.67-million-year-old Australopithecus specimen StW 573 ('Little Foot') at the I12 beamline of the Diamond Light Source (United Kingdom). Besides showing fine details of the enamel (i.e., hypoplasias) and cementum (i.e., incremental lines), as well as of the cranial bone microarchitecture (e.g., diploic channels), our synchrotron-based investigation reveals for the first time the 3D spatial organization of the Haversian systems in the mandibular symphysis of an early hominin.

Osteon circularity and longitudinal morphology: Quantitative and qualitative three-dimensional perspectives on human Haversian systems.

OBJECTIVES: Haversian systems result from bone remodeling, and show variation in size and shape among differing ages, body weights, mechanical environments, and species. While variables such as osteon circularity (On.Cr.) are generally studied in single transverse cross-sections, little is known about On.Cr. variation along an osteon's length, investigated here, in order to strengthen our understanding of bone microstructure. MATERIALS AND METHODS: Up to 875 measurements of On.Cr. were generated for 41 osteonal segments from the proximal anterior diaphysis of femoral human cortical bone of three adult male samples (ages 46, 62, 74). We employed four hypotheses to investigate On.Cr. variability, in cross-section and longitudinally. H1: There is no difference in On.Cr. among osteons comprising single cross-sections, H2: There is no difference in On.Cr. among individuals when single cross-sections are compared, H3: There is no difference in On.Cr. among measurements taken from an osteon along the longitudinal axis, and H4: There is no discernable pattern in an osteon's deviation from circularity. RESULTS: Quantitative analysis of single cross-sections revealed relatively consistent On.Cr. measurements within individual cross-sections and among individuals, supporting both, H1 and H2. Along individual osteonal segments, substantial degrees of dispersion from central tendencies were observed in 27 out of 41 analyzed osteons (despite relatively low overall standard deviations and interquartile ranges), leading to a rejection of H3. Qualitative characterization of morphological deviation from a "typical" circularity suggests a patterned deviation, leading also to a rejection of H4. DISCUSSION: On.Cr. variation is discussed in the context of both, phenomena intrinsic to a given osteon (including repetitive, small perturbations at roughly 45 µm intervals), and extrinsic (including shared reversal sheaths, osteonal branching, transverse connections, and osteonal repathing). Interesting associations between On.Cr. and other characteristics of the local Haversian network emphasize the role of Haversian systems as integrated parts of a greater morphological complex.

A first comparison of bone histomorphometry in extant domestic horses (Equus caballus) and a Pleistocene Indian wild horse (Equus namadicus).

The microstructural features of the tissue of long bones subjected to different biomechanical stresses could be a helpful tool for a better understanding of locomotor behavior in extant and extinct mammals, including equids. However, few researches have attempted to describe the bone tissue of extinct horses. In our study, we analyze and compare the histomorphometric features of the bone tissue in extant modern horses, Equus caballus, and Equus namadicus, a Pleistocene Indian extinct wild horse. The number, position, and size of the osteons and Haversian canals of the bone tissue, classifiable as dense Haversian tissue, were considered for the comparison. The results obtained highlight some differences between the analyzed species, E. caballus having fewer and bigger osteons than E. namadicus. The microstructural differences may depend on the different lifestyles and environmental conditions characterizing the two species. The results obtained suggest that comparing the biomechanical properties of extinct and modern horse species may provide indirect information on their paleoenvironment.

Three-dimensional topography of scapular nutrient foramina.

PURPOSE: The aim of this study is to describe the number and location of the nutrient foramina in human scapulae which can minimize blood loss during surgery. METHODS: 30 cadaveric scapulae were macerated to denude the skeletal tissue. The nutrient foramina of 0.51 mm and larger were identified and labeled by adhering glass beads. CT scans of these scapulae were segmented resulting in a surface model of each scapula and the location of the labeled nutrient foramina. All scapulae were scaled to the same size projecting the nutrient foramina onto one representative scapular model. RESULTS: Average number of nutrient foramina per scapula was 5.3 (0-10). The most common location was in the supraspinous fossa (29.7%). On the costal surface of the scapula, most nutrient foramina were found directly inferior to the suprascapular notch. On the posterior surface, the nutrient foramina were identified under the spine of the scapula in a somewhat similar fashion as those on the costal surface. Nutrient foramina were least present in the peri-glenoid area. CONCLUSION: Ninety percent of scapulae have more than one nutrient foramen. They are located in specific areas, on both the posterior and costal surface.

Morphometric analyses of clavicle's nutrient foramen.

PURPOSE: Fractures of the clavicle, which has an important location and function in the upper extremity and shoulder joint, compose 10% of all fracture cases. During the osteosynthesis of clavicle fractures and in the post-operative period of patients, considering the detailed morphometric and topographic properties of the nutrient foramen of clavicle is important to avoid the disruption of arterial nutrition of the clavicle and prevent unexpected injuries. The aim of this study was to investigate the morphometric properties of the nutrient foramen of clavicle in more detail using computedtomography images. METHODS: Computed tomography images of 116 healthy individuals (56 women/60 men) who had no pathology history were included in the presented study. Computed tomography images were reconstructed three-dimensionally using free-licensed Horos v3.3.3 software. Then, distances from clavicle's nutrient foramen to sternal end, anterior and posterior edges of the clavicle were measured. Statistical analyses were completed using SPSS v21 software. RESULTS: Our results demonstrated that the nutrient foramen of clavicle was located closer to the sternal end of the clavicle. The shortest distance to the sternal edge of clavicle was measured as 3.3 cm. Analyses of gender differences indicated that statistically significant differences were in favor of men. However, topographic properties of the clavicle's nutrient foramen were not affected by age. CONCLUSION: Nutrient foramen is mostly located closer to the sternal end of clavicle. Especially during osteosynthesis of clavicle fractures at the sternal end, maintaining the arterial supply of clavicle is of great importance for increasing the post-operative life quality of patients.

Inter-population variation of histomorphometric variables used in the estimation of age-at-death.

Population variation of several microscopic structures used in age-at-death estimation was assessed for three different population samples. The aim of the study was to determine if the need exists for population-specific standards when dealing with individuals of African and European origin. A total sample 223 bone sections from the anterior cortex of the femur (n = 99 black South Africans, n = 94 white South Africans and n = 30 Danish individuals) were analysed using a stereological protocol. Variables assessed included the average number of osteons per grid area (OPD), osteon size and Haversian canal size. ANCOVA was employed for assessment of statistically significant differences. The results indicated that OPD differed significantly between the three groups, but that osteon size was similar for all individuals. Haversian canal size showed unpredictable changes with age and high levels of variation, making it unsuitable to use for age estimation as a single factor. As there are conflicting opinions in the literature on whether to use population-specific equations for the estimation of age-at-death or not, this paper provided additional insight into the use of specific variables and its related variation between groups.

Relation between cross-sectional bone geometry and double zonal osteon frequency and morphology.

OBJECTIVES: While double-zonal osteons (DZ) are characterized by a hyper-mineralized ring inside their lamellae, recent findings suggest that this ring is also defined by a change in the collagen fibers' orientation. Collagen and minerals are essential components to the maintenance of adequate bone strength and their alteration can modify the mechanical properties of the bone tissue. Consequently, the aim of this study is to explore the effect of past loads, as estimated from cross-sectional geometric properties, on the formation of DZ osteons compared to type I (common) osteons. MATERIALS AND METHODS: The sample consists of paired humerus and femur midshaft sections (n = 23) of Eurocanadian settlers from the historical St. Matthew cemetery, Quebec City (1771-1860). Histomorphometric variables included in this study are osteon density for DZ and type I osteons (DZD; OPD), osteon area (DZOn.Ar; On. Ar), Haversian canal area (DZH.Ar; H.Ar), and the area within the hypermineralized ring (HR. Ar). Loading history is estimated from cross-sectional properties including the following variable: cortical and total area (CA, TA), maximum and minimum second moment of area (Imax , Imin ) and polar moment of area (J). RESULTS: When the humerus and femur of the same individuals are compared, the femur has a higher OPD, DZD, and relative DZD (DZD/OPD). DZ osteons have a smaller area and Haversian canal area compared to type I osteons. The area within the hypermineralized ring in DZ is higher than the Haversian canal area of the type I osteons. Correlations between the residual scores of the regression of histomorphometric variables and cross-sectional properties of the humerus on the femur were not significant. DISCUSSION: Based on the analysis of the entire cross-section, the lack of correlation between variations in cross-sectional properties and remodeling combined with the significant differences between humeri and femura suggests that the creation of DZ or type I osteons in the bone tissue might be due to a bone specific response, possibly related to differences in bone tissue age that needs to be further investigated. Definitive conclusion regarding biomechanical loads still seem to be premature as regional variations associated with mechanical properties remain to be explored.

What about limb long bone nutrient canal(s)? - a 3D investigation in mammals.

The nutrient arteries, located in the long bone diaphysis, are the major blood supply to long bones, especially during the early phases of growth and ossification. Their intersection with the central axis of the medullary area corresponds to the ossification center, and their opening on the outer bone surface to the nutrient foramen. Nutrient arteries/foramen have essentially been analyzed in humans, and only to a much lesser extent in a few mammals. Some studies have taken measurements of the nutrient foramen; others have investigated the shape and orientation of the nutrient canals, although only partially. No studies have analyzed the nutrient canal in three dimensions inside the bone and the relationships between nutrient foramen, nutrient canal, growth, and physiology require further investigation. The current study proposes to investigate in three dimensions the shape of the nutrient canal in stylopod bones of various mammals. Qualitative and quantitative parameters are defined to discuss the diversity in, for example, morphology, orientation, and diameter encountered, resorting to two different datasets to maximize differences within mammals and then analyze variation within morphologically and phylogenetically closer taxa. This study highlights a strong intraspecific variation for various parameters, with limited biological signal, but also shows trends. It notably provides evidence that canals are generally more numerous and relatively thinner in less elongated bones. Moreover, it shows that the growth center is located distally in the humerus and proximally in the femur, and that the canals are essentially oriented towards the faster growing end, so that the nutrient foramen does not indicate the location of the growth center. This result seems general in mammals but cannot be generalized outside of Mammalia. Further analyses of the features of nutrient arteries in reptiles are required to make comparisons with the trends observed in mammals.

Using nutrient foramina to differentiate human from non-human long bone fragments in bioarchaeology and forensic anthropology.

Long bone shaft fragments can be found isolated in archaeological and forensic contexts, such as in mass fatality incidents. When diagnostic morphological landmarks are not visible, the assessment of a human or non-human origin of a bone fragment can be challenging. Further methods need to be developed. In long bones, the presence of a nutrient foramen on fragments that do not show any diagnostic landmarks can make the assessment of the origin of the bone still possible. In this paper, human long bones were compared to those of the following species: chicken (Gallus gallus domesticus), duck (Cairina moschata), sheep (Ovis aries), pig (Sus scrofa domesticus), and deer (fallow deer, Dama dama, and roe deer, Capreolus capreolus). Macroscopic evaluation of location, direction and appearance, and micro-CT scanning in order to measure angle and shape of canal entrance, were applied in this study to differentiate human from non-human nutrient foramina on long bone shafts. Observations regarding the location and direction of nutrient foramina were proven to be different between human and non-human bones; however, these two features might not be exploitable in cases of highly fragmented bones. The foramina appearance was the most reliable for the origin identification, although the shape of the canal entrance and its angle at the cortical bone, obtained from micro-CT scans, were also useful parameters. For a correct identification of a fragment, one parameter may not be enough and it is advisable to employ as many features as possible. This research demonstrates that nutrient foramina have potential as a reliable bone feature for the distinction between human and non-human fragmented and incomplete long bones.

3D analysis of the osteonal and interstitial tissue in human radii cortical bone.

Human cortical bone has a complex hierarchical structure that is periodically remodelled throughout a lifetime. This microstructure dictates the mechanical response of the tissue under a critical load. If only some structural features, such as the different porosities observed in bone, are primarily studied, then investigations may not fully consider the osteonal systems in three-dimensions (3D). Currently, it is difficult to differentiate osteons from interstitial tissue using standard 3D characterization methods. Synchrotron radiation micro-computed tomography (SR-µCT) in the phase contrast mode is a promising method for the investigation of osteons. In the current study, SR-µCT imaging was performed on cortical bone samples harvested from eight human radii (female, 50-91 y.o.). The images were segmented to identify Haversian canals, osteocyte lacunae, micro-cracks, as well as osteons. The significant correlation between osteonal and Haversian canal volume fraction highlights the role of the canals as sites where bone remodelling is initiated. The results showed that osteocyte lacunae morphometric parameters depend on their distance to cement lines, strongly suggesting the evolution of biological activity from the beginning to the end of the remodelling process. Thus, the current study provides new data on 3D osteonal morphometric parameters and their relationships with other structural features in humans.

Bone histomorphometric measures of physical activity in children from medieval England.

OBJECTIVES: Histomorphometric studies show consistent links between physical activity patterns and the microstructure underlying the size and shape of bone. Here, we adopt a combined bone approach to explore variation in microstructure of ribs and humeri related to physical activity and historical records of manual labor in skeletal samples of children (n = 175) from medieval England. The humerus reflects greater biomechanically induced microstructural variation than the rib which is used here as a control. Variation in microstructure is sought between regions in England (Canterbury, York, Newcastle), and between high- and low-status children from Canterbury. MATERIALS AND METHODS: Thin-sections were prepared from the humerus or rib and features of bone remodeling were recorded using high-resolution microscopy and image analysis software. RESULTS: The density and size of secondary osteons in the humerus differed significantly in children from Canterbury when compared to those from York and Newcastle. Among the older children, secondary osteon circularity and diameter differed significantly between higher and lower status children. DISCUSSION: By applying bone remodeling principles to the histomorphometric data, we infer that medieval children in Canterbury engaged in less physically demanding activities than children from York or Newcastle. Within Canterbury, high-status and low-status children experienced similar biomechanical loading until around 7 years of age. After this age low-status children performed activities that resulted in more habitual loading on their arm bones than the high-status children. This inferred change in physical activity is consistent with historical textual evidence that describes children entering the work force at this age.

Collagen fiber orientation pattern, osteon morphology and distribution, and presence of laminar histology do not distinguish torsion from bending in bat and pigeon wing bones.

Bone can adapt to its habitual load history at various levels of its hierarchical structural and material organization. However, it is unclear how strongly a bone's structural characteristics (e.g. cross-sectional shape) are linked to microstructural characteristics (e.g. distributions of osteons and their vascular canals) or ultrastructural characteristics [e.g. patterns of predominant collagen fiber orientation (CFO)]. We compared the cross-sectional geometry, microstructure and ultrastructure of pigeon (Columba livia domestica) humeri, and third metacarpals (B3M) and humeri of a large bat (Pteropus poliocephalus). The pigeon humerus is habitually torsionally loaded, and has unremodeled ('primary') bone with vessels (secondary osteons are absent) and high 'laminarity' because a large majority of these vessels course circularly with respect to the bone's external surface. In vivo data show that the bat humerus is also habitually torsionally loaded; this contrasts with habitual single-plane bending of the B3M, where in vivo data show that it oscillates back and forth in the same direction. In contrast to pigeon humeri where laminar bone is present, the primary tissue of these bat bones is largely avascular, but secondary osteons are present and are usually in the deeper cortex. Nevertheless, the load history of humeri of both species is prevalent/predominant torsion, producing diffusely distributed shear stresses throughout the cross-section. We tested the hypothesis that despite microstructural/osteonal differences in these pigeon and bat bones, they will have similar characteristics at the ultrastructural level that adapt each bone for its load history. We postulate that predominant CFO is this characteristic. However, even though data reported in prior studies of bones of non-flying mammals suggest that CFO would show regional variations in accordance with the habitual 'tension regions' and 'compression regions' in the direction of unidirectional habitual bending, we hypothesized that alternating directions of bending within the same plane would obviate these regional/site-specific adaptations in the B3M. Similarly, but for other reasons, we did not expect regional variations in CFO in the habitually torsionally loaded bat and pigeon humeri because uniformly oblique-to-transverse CFO is the adaptation expected for the diffusely distributed shear stresses produced by torsion/multidirectional loads. We analyzed transverse sections from mid-diaphyses of adult bones for CFO, secondary osteon characteristics (size, shape and population density), cortical thickness in quadrants of the cortex, and additional measures of cross-sectional geometry, including the degree of circular shape that can help distinguish habitual torsion from bending. Results showed the expected lack of regional CFO differences in quasi-circular shaped, and torsionally loaded, pigeon and bat humeri. As expected, the B3M also lacked CFO variations between the opposing cortices along the plane of bending, and the quasi-elliptical cross-sectional shape and regional microstructural/osteonal variations expected for bending were not found. These findings in the B3M show that uniformity in CFO does not always reflect habitual torsional loads. Osteon morphology and distribution, and presence of laminar histology also do not distinguish torsion from bending in these bat and pigeon wing bones.

Microscopic markers of an infradian biorhythm in human juvenile ribs.

Recent studies have indicated that there may be an infradian systemic biorhythm that coordinates aspects of human hard tissue growth and influences adult body size. Here we investigate if evidence of this biorhythm retained in human teeth as the periodicity of Retzius lines (RP) corresponds with the microstructural growth of a non-weight bearing bone, the rib, in a sample of 50 human juvenile skeletons. Using static histomorphometric methods, the RP of one permanent tooth from each skeleton was calculated and combined with measures of bone remodeling in a rib from the same individual. Results provide the first evidence that the infradian biorhythm is linked to bone remodeling in children. Retzius periodicity was negatively correlated with relative osteon area (r = -0.563, p = 0.008) and positively related to Haversian canal area (r = 0.635, p = 0.002) and diameter (r = 0.671, p = 0.001) in children between the age of 8 to 12 years. There was also a negative correlation between RP and the relative cortical area of ribs (r = -0.500, p = 0.048). Relationships between bone remodeling and the biorhythm were much more variable in younger children. Results imply that as the biorhythm speeds up there is increased bone deposition during remodeling of the rib, leading to the larger osteonal lamellar bone areas and smaller Haversian canals in children between 8 and 12 years of age. Our results support the idea that there is an infradian biorhythm that coordinates aspects of human hard tissue growth.

Investigating interindividual variations in cortical bone quality: analysis of the morphotypes of secondary osteons and their population densities in the human femoral diaphysis.

Osteons are the primary sites of cortical bone lesions. However, many aspects of osteon microstructure remain poorly understood. This study aimed to explores interindividual differences in the osteon morphotype distributions in the human femoral diaphysis by evaluating the secondary osteon distributions in samples from human femurs. Two anonymized bone fragments from two modern Japanese femurs were examined. Twelve continuous transverse femoral diaphysis specimens were prepared from each fragment. Imaging examinations were conducted using a circularly polarized light microscope, and cross-sectional images were rendered using graphical synthesis software. Osteons in the images were identified as either bright-type osteons, dark-type osteons, or an others type. The two femurs were compared, and the secondary osteon morphotype distributions in different regions of their cross-sections were analyzed. When the two femurs were compared, significant differences in osteon density were observed in some regions and cross-sections. The dark-type osteon presence was strongest in the anterior and posterior regions of the femurs. The analytical method used in this study was found to be able to evaluate osteon microstructure. The results suggest that examining additional specimens and analyzing the biomechanical underpinnings of interindividual differences in osteon distribution patterns may help to improve our understanding of osteon microstructure.

Bilateral Asymmetry of Nutrient Foramen Position in Forearm Bones: Implications for its Use in Sorting Commingled Remains.

Commingling is frequently encountered in cases examined by the Committee on Missing Persons in Cyprus, and pair-matching is routinely used as a preliminary segregation method. Here we examine the degree of bilateral asymmetry in nutrient foramen position of the radius and ulna to determine whether nutrient foramen position may be useful in visual and/or ostetometric pair-matching. Differences in nutrient foramen position between left-right pairs from the same individual were compared with differences in nutrient foramen position between different individuals. Bilateral asymmetry in nutrient foramen position was found to be high, indicating that it is not a reliable trait for pair-matching. Bilateral asymmetry and between-person variation were found to differ between the two bones examined, suggesting that nutrient foramen position in other long bones should also be tested.

3D micro structural analysis of human cortical bone in paired femoral diaphysis, femoral neck and radial diaphysis.

Human bone is known to adapt to its mechanical environment in a living body. Both its architecture and microstructure may differ between weight-bearing and non-weight-bearing bones. The aim of the current study was to analyze in three dimensions, the morphology of the multi-scale porosities on human cortical bone at different locations. Eight paired femoral diaphyses, femoral necks, and radial diaphyses were imaged using Synchrotron Radiation µCT with a 0.7 µm isotropic voxel size. The spatial resolution facilitates the investigation of the multiscale porosities of cortical bone, from the osteonal canals system down to the osteocyte lacunar system. Our results showed significant differences in the microstructural properties, regarding both osteonal canals and osteocytes lacunae, between the different anatomical locations. The radius presents significantly lower osteonal canal volume fraction and smaller osteonal canals than the femoral diaphysis or neck. Osteocytes lacunae observed in the radius are significantly different in shape than in the femur, and lacunar density is higher in the femoral neck. These results show that the radius, a non-weight-bearing bone, is significantly different in terms of its microstructure from a weight-bearing bone such as the femur. This implies that the cortical bone properties evaluated on the femoral diaphysis, the main location studied within the literature, cannot be generalized to other anatomical locations.

Brief communication: Test of a method to identify double-zonal osteon in polarized light microscopy.

OBJECTIVES: Double-zonal osteons (DZ) have been of interest in paleopathological research because they might be linked to physiological pathology. DZ are thought to be evidence of arrested osteon formation with a brief but abrupt increase in mineralization of lamellae occurring during bone remodeling. Originally identified from microradiographs as hypermineralized rings, recent studies have identified DZ from linear polarized light microscopy (PLM). However, PLM does not guarantee the adequate detection of DZ since PLM captures bone birefringence and not hyper-mineralization. Scanning electron microscopy with backscatter electrons (BSE-SEM) allows observation of DZ by detecting differences in mineralization. The purpose of this study is to investigate whether DZ, as identified by BSE-SEM, can indeed be identified with PLM. MATERIALS AND METHODS: The sample consists of an archaeological collection of adult midshaft femurs (n = 30) from St. Matthew cemetery, Quebec City (1771-1860). DZ were identified and counted independently with PLM and BSE-SEM for the same sections. Results from both methods were compared. RESULTS: Chi-square test shows that there was no significant difference between the two methods (p = 0.404). No significant bias was found on Bland-Altman analysis and Cohen's kappa shows a substantial agreement between the two methods (Κ = 0.66). PLM shows a good accuracy (sensitivity 79%, specificity 99.4%) and reliability (Positive Predictive Value: 86.71%; Negative Predictive Value: 99.45%). DISCUSSION: These findings indicate that the two methods are interchangeable. PLM, using our proposed protocol, is reliable to accurately identify DZ. We discuss how PLM and BSE-SEM that measure different features of the bone tissue can converge on the identification of DZ.